Interactions of hydrolyzed β-lactams with the L1 metallo-β-lactamase: crystallography supports stereoselective binding of cephem/carbapenem products

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dc.citation.titleJournal of Biological Chemistry
dc.citation.volume299
dc.contributor.orcidhttps://orcid.org/0000-0002-1380-9824
dc.contributor.orcidhttps://orcid.org/0000-0002-7978-3233
dc.creatorHinchliffe, Philip
dc.creatorCalvopiña, Karina
dc.creatorRabe, Patrick
dc.creatorMojica, María F.
dc.creatorSchofield, Christopher J.
dc.creatorDmitrienko, Gary I.
dc.creatorBonomo, Robert A.
dc.creatorVila, Alejandro J.
dc.creatorSpencer, James
dc.date.accessioned2024-08-22T19:19:50Z
dc.date.available2024-08-22T19:19:50Z
dc.date.issued2023-03-15
dc.description.abstractL1 is a dizinc subclass B3 metallo-β-lactamase (MBL) that hydrolyzes most β-lactam antibiotics and is a key resistance determinant in the Gram-negative pathogen Stenotrophomonas maltophilia, an important cause of nosocomial infections in immunocompromised patients. L1 is not usefully inhibited by MBL inhibitors in clinical trials, underlying the need for further studies on L1 structure and mechanism. We describe kinetic studies and crystal structures of L1 in complex with hydrolyzed β-lactams from the penam (mecillinam), cephem (cefoxitin/cefmetazole), and carbapenem (tebipenem, doripenem, and panipenem) classes. Despite differences in their structures, all the β-lactam-derived products hydrogen bond to Tyr33, Ser221, and Ser225 and are stabilized by interactions with a conserved hydrophobic pocket. The carbapenem products were modeled as Δ1-imines, with (2S)-stereochemistry. Their binding mode is determined by the presence of a 1β-methyl substituent: the Zn-bridging hydroxide either interacts with the C-6 hydroxyethyl group (1β-hydrogen-containing carbapenems) or is displaced by the C-6 carboxylate (1β-methyl-containing carbapenems). Unexpectedly, the mecillinam product is a rearranged N-formyl amide rather than penicilloic acid, with the N-formyl oxygen interacting with the Zn-bridging hydroxide. NMR studies imply mecillinam rearrangement can occur nonenzymatically in solution. Cephem-derived imine products are bound with (3R)-stereochemistry and retain their 3′ leaving groups, likely representing stable endpoints, rather than intermediates, in MBL-catalyzed hydrolysis. Our structures show preferential complex formation by carbapenem- and cephem-derived species protonated on the equivalent (β) faces and so identify interactions that stabilize diverse hydrolyzed antibiotics. These results may be exploited in developing antibiotics, and β-lactamase inhibitors, that form long-lasting complexes with dizinc MBLs.
dc.description.filFil: Hinchliffe, Philip. University of Bristol, Biomedical Sciences Building. School of Cellular and Molecular Medicine; United Kingdom.
dc.description.filFil: Calvopiña, Karina. University of Oxford. Ineos Oxford Institute for Antimicrobial Research. Chemistry Research Laboratory. Department of Chemistry; United Kingdom.
dc.description.filFil: Rabe, Patrick. University of Oxford. Ineos Oxford Institute for Antimicrobial Research. Chemistry Research Laboratory. Department of Chemistry; United Kingdom.
dc.description.filFil: Mojica, María F. Case Western Reserve University. School of Medicine. Department of Molecular Biology and Microbiology; USA.
dc.description.filFil: Mojica, María F. CWRU-Cleveland VA Medical Center for Antimicrobial Resistance and Epidemiology. U.S. Department of Veterans Affairs; USA.
dc.description.filFil: Mojica, María F. Louis Stokes Cleveland. Research Service. Department of Veterans Affairs Medical Center; USA.
dc.description.filFil: Mojica, María F. Universidad El Bosque. Grupo de Resistencia Antimicrobiana y Epidemiología Hospitalaria; Colombia.
dc.description.filFil: Schofield, Christopher J. University of Oxford. Ineos Oxford Institute for Antimicrobial Research. Chemistry Research Laboratory. Department of Chemistry; United Kingdom.
dc.description.filFil: Dmitrienko, Gary I. University of Waterloo. Department of Chemistry; Canada.
dc.description.filFil: Dmitrienko, Gary I. University of Waterloo. School of Pharmacy: Canada.
dc.description.filFil: Bonomo, Robert A. CWRU-Cleveland VA Medical Center for Antimicrobial Resistance and Epidemiology. U.S. Department of Veterans Affairs; USA.
dc.description.filFil: Bonomo, Robert A. Louis Stokes Cleveland. Research Service. Department of Veterans Affairs Medical Center; USA.
dc.description.filFil: Bonomo, Robert A. Case Western Reserve University School of Medicine. Departments of Medicine, Biochemistry, Pharmacology, and Proteomics and Bioinformatics; USA.
dc.description.filFil: Vila, Alejandro J. CWRU-Cleveland VA Medical Center for Antimicrobial Resistance and Epidemiology. U.S. Department of Veterans Affairs; USA.
dc.description.filFil: Vila, Alejandro J. Instituto de Biología Molecular y Celular de Rosario. Laboratorio de Metaloproteínas (IBR-CONICET); Argentina.
dc.description.filFil: Vila, Alejandro J. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Área Biofísica; Argentina.
dc.description.filFil: Spencer, James. University of Bristol, Biomedical Sciences Building. School of Cellular and Molecular Medicine; United Kingdom.
dc.description.sponsorshipNational Institute of Allergy and Infectious Diseases of the National Institutes of Health: R01AI100560, R21-AI123835
dc.description.sponsorshipCanadian Institutes of Health Research: PJT427834
dc.description.sponsorshipBristol Centre for Synthetic Biolog
dc.format.extent1-15
dc.identifier.issn1083-351X
dc.identifier.issn0021-9258
dc.identifier.urihttps://hdl.handle.net/2133/27632
dc.language.isoen
dc.publisherElsevier
dc.publisherAmerican Society for Biochemistry and Molecular Biology
dc.relation.publisherversionhttps://doi.org/10.1016/j.jbc.2023.104606
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S002192582300248X?via%3Dihub
dc.rightsopenAccess
dc.rights.holderHinchliffe, Philip
dc.rights.holderCalvopiña, Karina
dc.rights.holderRabe, Patrick
dc.rights.holderMojica, María F.
dc.rights.holderSchofield, Christopher J.
dc.rights.holderDmitrienko, Gary I.
dc.rights.holderBonomo, Robert A.
dc.rights.holderVila, Alejandro J.
dc.rights.holderSpencer, James
dc.rights.holderUniversidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas
dc.rights.textAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectMetallo-β-lactamases
dc.subjectAntibiotic resistance
dc.subjectβ-lactam
dc.subjectCarbapenem
dc.subjectHydrolysis
dc.subjectX-ray crystallography
dc.subjectL1 carbapenemase
dc.titleInteractions of hydrolyzed β-lactams with the L1 metallo-β-lactamase: crystallography supports stereoselective binding of cephem/carbapenem products
dc.typearticulo
dc.type.collectionarticulo
dc.type.versionpublishedVersion

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